Rapamycin Improves Palmitate-Induced ER Stress/NF κ B Pathways Associated with Stimulating Autophagy in Adipocytes

Figure 2

Autophagy is activated in response to palmitate and rapamycin. (a) Adipocytes were treated with or without palmitate (0.5 or 1.0 mM) and rapamycin (50 or 100 nM) for 12 h, followed by LC3 Western blotting analysis. (b) Preincubated with or without CQ (10 μM) for 1 h and with BSA or BSA-conjugated PA for 12 h, followed by LC3 Western blotting analysis. (c) Electron microscopy and MDC staining of mature adipocytes expose to 0.5 mM PA or 100 nM RAP for 12 h. Lipid droplets (white arrow); autophagosomes and autolysosome of double-membrane, single-membrane, and multivesicular body-like vesicles (blue arrows); endoplasmic reticulum (red arrow); mitochondria (yellow arrow). Scale bars are indicated at bottom. (d) Pretreat with or without 4-PBA (10 mM) for 1 h and with PA (0.5 mM) for 12 h followed by LC3 Western blotting analysis. The proteins are quantified and normalized to GAPDH. Results are mean ± SEM of three to five separate experiments. versus nontreatment group (0.5% BSA); ^# versus PA 0.5 mM group.

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